Pharmaceutical composition for in vitro amplification of cytotoxic gamma delta T-lymphocytes and amplification method of pharmaceutical composition

Abstract

The invention relates to the field of immunology, in particular to pharmaceutical composition for in vitro amplification of gamma delta T cells and an amplification method of the pharmaceutical composition and provides pharmaceutical composition for in vitro amplification of cytotoxic gamma delta T-lymphocytes and an amplification method of the pharmaceutical composition to overcome the defects ofcomplexity of existing methods for acquiring gamma delta T cells. The pharmaceutical composition comprises ZOL (zoledronate) with the concentration of 1-5 mu mol / L and POM (pomalidomide) with the concentration of 0.1-1.2 mu mol / L. The provided POM and ZOL pharmaceutical composition can realize proliferation and activation of cytotoxic gamma delta T cells. Compared with existing methods for acquiring gamma delta T cells, the provided amplification method comprises simple procedures, injection of IL-2 is not required, the clinical applicability is high and the cost is lower.

Description

technical field

[0001] The present invention relates to the field of immunology. Specifically, the present invention relates to a drug combination for expanding cytotoxic γδT cells in vitro and an expansion method thereof. Background technique

[0002] Human peripheral blood γδT cells are a subset of CD3-positive T lymphocytes, accounting for about 5-10% of peripheral blood lymphocytes.

[0003] Previous studies have shown that the drug zoledronate, which has been approved for the clinical treatment of osteoporosis, can activate and expand human peripheral blood γδT cells under the condition of cytokine participation, so that the proportion of cytotoxic γδT cells in them increases sharply High, cytotoxic γδT cells can target and kill a variety of tumor cells in vitro, and have good prospects for clinical application. At present, the preparation of cytotoxic γδT cells in vitro is mainly achieved by co-culturing human peripheral blood mononuclear cells with zoledronic acid a...

Examples

Embodiment 1

[0077] Take the peripheral blood of the patient whose number (#) is 1, and amplify according to the amplification method provided by the present invention. The result is as figure 1 shown. The data on the proportion of cytotoxic γδ T cells are entered in Table 1.

[0078] Among them, the concentration of ZOL is 1umol / L, the activity of IL-2 is 100U / mL, and the concentration of POM is 1umol / L.

[0079] Control is a blank control, that is, no drugs are added, only cell culture fluid and cells are added. Zol, IL-2, and POM were single-drug controls, and the rest were drug combinations.

[0080] figure 1 , the value in the upper right quadrant of each figure represents the proportion of cytotoxic γδT cells, and the larger the value in the upper right quadrant, the higher the proportion of cytotoxic γδT cells in the culture system and the greater the number of cells.

[0081] According to the detection principle of flow cytometry, the values ​​in each quadrant represent differ...

Embodiment 2

[0083] Take the peripheral blood of the healthy person whose number (#) is 8, and amplify according to the amplification method provided by the present invention. The result is as figure 2 shown. The data on the proportion of cytotoxic γδ T cells are entered in Table 1.

Embodiment 3

[0085] Take the peripheral blood of healthy people whose numbers (#) are 8, 11, 12, 15-17, and amplify according to the amplification method provided by the present invention. The result is as image 3 As shown, the amplification effect of the drug combination of POM and ZOL provided by the present invention is equivalent to or better than that of IL-2 and ZOL. The data on the proportion of cytotoxic γδ T cells is entered in Table 2.

[0086] Table 1 The proportion of expanded cytotoxic γδT cells in Examples 1-2 provided by the present invention

[0087]

[0088] Table 2 The proportion of expanded cytotoxic γδT cells in Example 3 provided by the present invention

[0089]

technical field

[0001] The present invention relates to the field of immunology. Specifically, the present invention relates to a drug combination for expanding cytotoxic γδT cells in vitro and an expansion method thereof. Background technique

[0002] Human peripheral blood γδT cells are a subset of CD3-positive T lymphocytes, accounting for about 5-10% of peripheral blood lymphocytes.

[0003] Previous studies have shown that the drug zoledronate, which has been approved for the clinical treatment of osteoporosis, can activate and expand human peripheral blood γδT cells under the condition of cytokine participation, so that the proportion of cytotoxic γδT cells in them increases sharply High, cytotoxic γδT cells can target and kill a variety of tumor cells in vitro, and have good prospects for clinical application. At present, the preparation of cytotoxic γδT cells in vitro is mainly achieved by co-culturing human peripheral blood mononuclear cells with zoledronic acid a...

Examples

Embodiment 1

[0077] Take the peripheral blood of the patient whose number (#) is 1, and amplify according to the amplification method provided by the present invention. The result is as figure 1 shown. The data on the proportion of cytotoxic γδ T cells are entered in Table 1.

[0078] Among them, the concentration of ZOL is 1umol / L, the activity of IL-2 is 100U / mL, and the concentration of POM is 1umol / L.

[0079] Control is a blank control, that is, no drugs are added, only cell culture fluid and cells are added. Zol, IL-2, and POM were single-drug controls, and the rest were drug combinations.

[0080] figure 1 , the value in the upper right quadrant of each figure represents the proportion of cytotoxic γδT cells, and the larger the value in the upper right quadrant, the higher the proportion of cytotoxic γδT cells in the culture system and the greater the number of cells.

[0081] According to the detection principle of flow cytometry, the values ​​in each quadrant represent differ...

Embodiment 2

[0083] Take the peripheral blood of the healthy person whose number (#) is 8, and amplify according to the amplification method provided by the present invention. The result is as figure 2 shown. The data on the proportion of cytotoxic γδ T cells are entered in Table 1.

Embodiment 3

[0085] Take the peripheral blood of healthy people whose numbers (#) are 8, 11, 12, 15-17, and amplify according to the amplification method provided by the present invention. The result is as image 3 As shown, the amplification effect of the drug combination of POM and ZOL provided by the present invention is equivalent to or better than that of IL-2 and ZOL. The data on the proportion of cytotoxic γδ T cells is entered in Table 2.

[0086] Table 1 The proportion of expanded cytotoxic γδT cells in Examples 1-2 provided by the present invention

[0087]

[0088] Table 2 The proportion of expanded cytotoxic γδT cells in Example 3 provided by the present invention

[0089]

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